Great job! If you were able to re-do the experiment with an accurate loading control and real time PCR, how would you think your results would change? Do you think that it would support your hypothesis?
I don’t know that our results would change that much, because the expression for Cory is so low even after DNA damage was induced. If the experiment was repeated and we got a proper loading control we would just know how much more intense that band is. Again, it is not very much. The consensus gel had much the same as what we got in terms of Cory expression. Real time PCR would just let us know exactly how much of the mRNA was produced after DNA damage occurred.
Real time PCR is just a way of measuring exactly how much mRNA is produced after the DNA damage occurred. So rather than a vague “this band is brighter than that” we have a pretty accurate and quantitative way of knowing how much of the mRNA was produced, which is a way to measure gene expression.
We think that the primers were annealing to themselves more than they were to the gene. In addition, the expression for Cory is so faint even after DNA damage that it is possible that the it isn’t expressed during normal cellular function (sans DNA damage). So that means that there wouldn’t be any cDNA for the primers to anneal to in the first place.
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Great job! If you were able to re-do the experiment with an accurate loading control and real time PCR, how would you think your results would change? Do you think that it would support your hypothesis?
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I don’t know that our results would change that much, because the expression for Cory is so low even after DNA damage was induced. If the experiment was repeated and we got a proper loading control we would just know how much more intense that band is. Again, it is not very much. The consensus gel had much the same as what we got in terms of Cory expression. Real time PCR would just let us know exactly how much of the mRNA was produced after DNA damage occurred.
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Referring to your future directions, how would using real time PCR compare to the PCR you did use in the experiment?
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Real time PCR is just a way of measuring exactly how much mRNA is produced after the DNA damage occurred. So rather than a vague “this band is brighter than that” we have a pretty accurate and quantitative way of knowing how much of the mRNA was produced, which is a way to measure gene expression.
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Why do you think the primers didn’t anneal? Was it because of a lack of info, something wrong with procedure, or plain bad luck?
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We think that the primers were annealing to themselves more than they were to the gene. In addition, the expression for Cory is so faint even after DNA damage that it is possible that the it isn’t expressed during normal cellular function (sans DNA damage). So that means that there wouldn’t be any cDNA for the primers to anneal to in the first place.
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